首页|Study on the preservation effects of the amputated forelimb by machine perfusion at physiological temperature

Study on the preservation effects of the amputated forelimb by machine perfusion at physiological temperature

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Purpose:Ischemia and hypoxia are the main factors limiting limb replantation and transplantation.Static cold storage(SCS),a common preservation method for tissues and organs,can only prolong limb ischemia time to 4-6 h.The normothermic machine perfusion(NMP)is a promising method for the preservation of tissues and organs,which can extend the preservation time in vitro by providing continuous oxygen and nutrients.This study aimed to evaluate the difference in the efficacy of the 2 limb preservation methods.Methods:The 6 forelimbs from beagle dogs were divided into 2 groups.In the SCS group(n=3),the limbs were preserved in a sterile refrigerator at 4 ℃ for 24 h,and in the NMP group(n=3),the perfusate prepared with autologous blood was used for the oxygenated machine perfusion at physiological tem-perature for 24 h,and the solution was changed every 6 h.The effects of limb storage were evaluated by weight gain,perfusate biochemical analysis,enzyme-linked immunosorbent assay,and histological analysis.All statistical analyses and graphs were performed using GraphPad Prism 9.0 one-way or two-way analysis of variance.The p value of less than 0.05 was considered to indicate statistical significance.Results:In the NMP group,the weight gained percentage was 11.72％±4.06％;the hypoxia-inducible factor-1α contents showed no significant changes;the shape of muscle fibers was normal;the gap be-tween muscle fibers slightly increased,showing the intercellular distance of(30.19±2.83)pm;and the vascular α-smooth muscle actin(α-SMA)contents were lower than those in the normal blood vessels.The creatine kinase level in the perfusate of the NMP group increased from the beginning of perfusion,decreased after each perfusate change,and remained stable at the end of perfusion showing a peak level of 4097.6 U/L.The lactate dehydrogenase level of the NMP group increased near the end of perfusion and reached the peak level of 374.4 U/L.In the SCS group,the percentage of weight gain was 0.18％±0.10％,and the contents of hypoxia-inducible factor-1α increased gradually and reached the maximum level of(164.85±20.75)pg/mL at the end of the experiment.The muscle fibers lost their normal shape and the gap between muscle fibers increased,showing an intercellular distance of(41.66±5.38)pm.The con-tents of vascular α-SMA were much lower in the SCS group as compared to normal blood vessels.Conclusions:NMP caused lesser muscle damage and contained more vascular α-SMA as compared to SCS.This study demonstrated that NMP of the amputated limb with perfusate solution based on autologous blood could maintain the physiological activities of the limb for at least 24 h.

Traumatic amputationMachine perfusionStatic cold storageLimb replantation

Sheng-Feng Chen、Bo-Yao Yang、Tie-Yuan Zhang、Xiang-Yu Song、Zhi-Bo Jia、Lei-Jia Chen、Meng-Yi Cui、Wen-Jing Xu、Jiang Peng

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Guizhou Medical University,Guiyang,550025,China

Medical School of PLA,Beijing,100048,China

Beijing Key Lab of Regenerative Medicine in Orthopedics,Key Laboratory of Musculoskeletal Trauma and War Injuries PLA,Department of Orthopedics,The Fourth Medical Center,Chinese PLA General Hospital,Beijing,100048,China

Hebei North University,Zhangjiakou,075000,Hebei Province,China

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国家重点研发计划Military Logistic(s)Scientific Research Program of China

2019YFA0110704BHJ20J002

2024

10.1016/j.cjtee.2023.05.005

中华创伤杂志(英文版)
中华医学会

中华创伤杂志(英文版)

CSTPCD
影响因子：0.608
ISSN：1008-1275
年,卷(期)：2024.27(2)
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